Measurement of the concentration of the haemoglobin and determination of the leukocytes in particular of specific leukocytic sub-populations, is of primordial importance in diagnosis of specific pathologies, both in human and veterinary medicine.
Haemoglobin is a chromo-protein which is contained in the red globules (or erythrocytes) of the blood.
Measurement of the concentration of the haemoglobin thus requires use of a cellular lysis reagent, which can carry out lysis of the erythrocytes or red globules, in order to release the haemoglobin, so that the latter can be measured.
For this purpose, it is known to use reagents which contain cyanide ions, which can carry out lysis of the erythrocyte, and transform the haemoglobin into a stable chromogen compound, in order to permit determination of the haemoglobin by means of colorimetry measurement. A reagent of this type is described in particular in U.S. Pat. No. 3,874,852. The main disadvantage of this reagent is that it uses cyanide. In addition, it does not make it possible to identify and quantify the leukocyte sub-populations which are contained in the blood sample to be analysed.
In fact, the determination of leukocyte sub-populations, in particular of lymphocytes, monocytes, neutrophils, and eosinophils, is of primordial importance for identification of specific pathologies.
Various reagents which do not contain cyanide have already been proposed in the prior art, for determination of the leukocytes in a blood sample.
Examples of reagents of this type are described for example in document EP 0 325 710.
Although these reagents have the advantage that they do not use cyanide, they have the disadvantage that they do not permit accurate identification and quantification of the leukocyte sub-populations of the blood.
Thus, the reagent according to EP 0 325 710 makes it possible, for example, to determine the eosinophils which are present in a blood sample, without however making it possible to identify accurately each of the other leukocyte sub-populations.
In addition, document EP 0 424 871 describes reagents which permit differentiation of leukocyte sub-populations. However, these reagents sometimes encounter difficulties in overcoming the membranous resistance which is present in human pathology and in veterinary biology.
The object of the invention is in particular to eliminate the aforementioned disadvantages.
In particular, it aims to provide a haematological analysis reagent for measurement of the haemoglobin and determination of the leukocytes in a blood sample, which in particular permits lysis of the erythrocytes or red globules, as well as measurement of the haemoglobin, without using cyanide compounds.
The object of the invention is also to provide a reagent of this type, which makes it possible to quantify the total leukocytes or white globules, to identify at least one leukocyte sub-population, and to quantify at least one leukocyte sub-population.
The object of the invention is also to provide a reagent of this type which can be used both in human and veterinary medicine.
In addition, the object of the invention is to provide a reagent of this type which can eliminate the analysis problems which are associated with the membranous resistance which is encountered in human pathology and in veterinary biology.